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Innovative High Gas Pressure Microscopy Chamber Designed for Biological Cell Observation

Published online by Cambridge University Press:  26 January 2016

Mélanie Ragon
Affiliation:
UMR A 02.102 Procédés Alimentaires et Microbiologiques, Université Bourgogne Franche-Comté/AgroSup Dijon, 1 Esplanade Erasme, 21000 Dijon, France
Hue Nguyen Thi Minh
Affiliation:
UMR A 02.102 Procédés Alimentaires et Microbiologiques, Université Bourgogne Franche-Comté/AgroSup Dijon, 1 Esplanade Erasme, 21000 Dijon, France
Stéphane Guyot
Affiliation:
UMR A 02.102 Procédés Alimentaires et Microbiologiques, Université Bourgogne Franche-Comté/AgroSup Dijon, 1 Esplanade Erasme, 21000 Dijon, France
Pauline Loison
Affiliation:
UMR A 02.102 Procédés Alimentaires et Microbiologiques, Université Bourgogne Franche-Comté/AgroSup Dijon, 1 Esplanade Erasme, 21000 Dijon, France
Gaëtan Burgaud
Affiliation:
Laboratoire Universitaire de Biodiversité et Ecologie Microbienne (EA3882), IFR 148, Université Européenne de Bretagne/Université de Brest/ESMISAB, Technopole Brest-Iroise, 29280 Plouzané, France
Sébastien Dupont
Affiliation:
UMR A 02.102 Procédés Alimentaires et Microbiologiques, Université Bourgogne Franche-Comté/AgroSup Dijon, 1 Esplanade Erasme, 21000 Dijon, France
Laurent Beney
Affiliation:
UMR A 02.102 Procédés Alimentaires et Microbiologiques, Université Bourgogne Franche-Comté/AgroSup Dijon, 1 Esplanade Erasme, 21000 Dijon, France
Patrick Gervais
Affiliation:
UMR A 02.102 Procédés Alimentaires et Microbiologiques, Université Bourgogne Franche-Comté/AgroSup Dijon, 1 Esplanade Erasme, 21000 Dijon, France
Jean-Marie Perrier-Cornet*
Affiliation:
UMR A 02.102 Procédés Alimentaires et Microbiologiques, Université Bourgogne Franche-Comté/AgroSup Dijon, 1 Esplanade Erasme, 21000 Dijon, France
*
*Corresponding author. [email protected]
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Abstract

An original high-pressure microscopy chamber has been designed for real-time visualization of biological cell growth during high isostatic (gas or liquid) pressure treatments up to 200 MPa. This new system is highly flexible allowing cell visualization under a wide range of pressure levels as the thickness and the material of the observation window can be easily adapted. Moreover, the design of the observation area allows different microscope objectives to be used as close as possible to the observation window. This chamber can also be temperature controlled. In this study, the resistance and optical properties of this new high-pressure chamber have been tested and characterized. The use of this new chamber was illustrated by a real-time study of the growth of two different yeast strains – Saccharomyces cerevisiae and Candida viswanathii – under high isostatic gas pressure (30 or 20 MPa, respectively). Using image analysis software, we determined the evolution of the area of colonies as a function of time, and thus calculated colony expansion rates.

Type
Biological Applications
Copyright
© Microscopy Society of America 2016 

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